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ANR funded project

Enjeux agroalimentaires (DS0502)
Edition 2014


MUcosal salivary Film and Flavor INteractions

MUcosal salivary Film & Flavor Interactions
MUFFIN proposes to elucidate the molecular basis of interactions between food constituents and the mucosal pellicle, focusing on two sensory modalities: astringency and aroma perception.

Objectives of MUFFIN
The objectives of MUFFIN is to test the following hypotheses:
1. the presence of MUC1 facilitates the anchoring of salivary proteins on mucosa and is thereby a facilitator of mucosal
pellicle formation
2. the mucosal pellicle plays a role in aroma persistence in the oral cavity;
3. the salivary PRPs protect the mucosal pellicle against the effects of tannins;
4. the effects of tannins on the mucosal pellicle modify its ability to interact with aroma compounds;
5. the inter-individual variations of the mucosal pellicle or saliva composition modify the interactions with flavor

Methods used in MUFFIN
MUFFIN used the following methods:
• for the WP1: cell culture, an optical microscopy and atomic force microscopy.
• for the WP2: GC-FID and PTR-MS
• for the WP3: optical microscopy and atomic force microscopy.


During the last 18 months, research were conducted on WP1, WP2 and WP3.
The main objective of WP1 is to establish a model of oral mucosa.
A model of oral mucosa has been developed using the cell line TR146 transfected by the gene MUC1. Immunostaining experiments have demonstrated the anchoring of the salivary mucin MUC5B at the surface of the cell, demonstrating the formation of the mucosal pellicle on the model of mucosa.
In the WP2, in order to show the phenomena of aromatic persistence and the adsorption of aroma compounds at the surface of the mucosa, experiments of aroma release have been performed. A first study on a linear serie of methyl ketones have been realized. These experiments have not shown an adsorption of the aroma compounds studied onto the mucosa. However, they have revealed a metabolization of the 2-nonanone and the 2-decanone by the cells of the mucosa.


The model of mucosa developed in the WP1 will be used to study the effects of tannins on the structure of the mucosal pellicle in order to have a better inter standing of the molecular phenomena involved in astringency. The WP4 will focus on the effect of tannins in aroma persistence, while the WP5 will study the effect of inter-individual variation on both astringency and aroma persistence.

Scientific outputs and patents


Ployon S, Belloir C, Bonnotte A, Lherminier J, Canon F & Morzel M. 2016. The membrane-associated MUC1 improves adhesion of salivary MUC5B on buccal cells. Application to development of an in vitro cellular model of oral epithelium. Archives of Oral Biology, 61:149-155.


Ployon S, Sémon E, Le Quéré J-L, Belloir C, Tromelin A, Guichard E, Morzel M & Canon F. 2015. Développement d’une méthode d’analyse de la désorption de composés d’arôme par PTR-MS sur un modèle in vitro de muqueuse orale. Congrès français de Spectrométrie de Masse et d'Analyse Protéomique, Ajaccio, 15-18 Septembre 2015.

Aybeke EN, Ployon S, Brulé, Bourillot E, Morzel M, Lesniewska E & Canon F. 2016. Impact of a mucosal pellicle on the physical properties of oral epithelial cells. International Scanning Probe Microscopy Conference, Grindelwald, Switzerland,12th-15th June 2016.

Ployon S, Pradels C, Belloir C, Morzel M & Canon F. 2016. Role of oral mucosa and mucosal pellicle in aroma persistence. 94th IADR (International Association for Dental Research) General Session, Seoul, Korea, 22nd-25th June 2016.

Oral communication

Ployon S, Pradels C, Belloir C, Morzel M & Canon F. 2016. Interactions between aroma compounds and the oral mucosa could be responsible for aroma persistence. 4th International Conference on Food Oral Processing (FOP), Lausanne, Switzerland, 3rd-6th July 2016.


CSGA Centre des Sciences du Goût et de l'Alimentation

ANR grant: 398 662 euros
Beginning and duration: octobre 2014 - 48 mois

Submission abstract

Flavor perception is one of the factors guiding food consumption. Flavor is mainly perceived by the chemical senses of taste and olfaction (aroma retro-nasal perception), but also by the “trigeminal senses”, which can detect for example astringent molecules. The effect of saliva and salivary proteins on flavor perception is increasingly recognized. More specifically, recent studies suggest that the mucosal pellicle, a layer of salivary proteins bound to oral mucosa, could be involved in flavor perception. For example, a study on astringency concluded that salivary proteins in solution inhibit the mouthfeeling of astringency, which is mediated, at least in part, by salivary proteins adhered to buccal cells. However, the molecular interactions occurring between the proteins of the mucosal pellicle and flavor compounds are still not thoroughly understood and studies dedicated to that topic are lacking. MUFFIN proposes to elucidate the molecular basis of interactions between food constituents and the mucosal pellicle, focusing on two sensory modalities: astringency and aroma perception.
Astringency is defined as a complex sensation due to shrinking, drawing, or puckering of the epithelium, resulting from exposure to substances such as tannins. It is generally accepted that the interactions occurring between salivary proline-rich proteins (PRP) and tannins play a role in this sensation. The mechanism could involve interactions between the PRP•tannin complexes, after aggregation, and the proteins belonging to the mucosal pellicle. Another hypothesis postulates that PRP have a protective role toward tannins, decreasing the astringency sensation by protecting the mucosa from tannins.
The oral mucosa may also be involved in persistence of aroma in the mouth, a sensory phenomenon for which no mechanistic explanation is available. It has been reported that free salivary proteins, such as mucin and a-amylase, can affect the release of aroma compounds. However, salivary proteins in solution are swallowed at each deglutition. Therefore, we propose that the aroma persistence in the mouth could be due to molecular interactions between the bound proteins of the mucosal pellicle and aroma compounds.
MUFFIN proposes to develop a model mucosa allowing in vitro experiments to gain a deeper understanding of these molecular interactions. This basic project will bring crucial information on the role of the mucosal pellicle in flavor perception. More particularly, MUFFIN should provide a unique opportunity (1) to develop a well characterized model of mucosa composed of a realistic salivary protein layer anchoring onto an epithelium; (2) to determine the role of the mucosal pellicle into aroma release and persistence in mouth; (3) to characterize the impact of tannins on the structure of the salivary film, the role of the salivary PRPs in the protection of the mucosa; (4) to characterize the effect of tannins on the ability of the mucosal pellicle to interact with aroma compounds and (5), finally, to investigate the effects of inter-individual variations in the composition and the structure of the mucosal pellicle on its interactions with flavor molecules. This ambitious project takes into account two different modalities of flavor (astringency, aromatic perception) and their interaction, also the inter-individual physiological and sensory variability. Therefore, it will provide a global vision of the role played by the mucosal pellicle on flavor.
To achieve its objectives, MUFFIN proposes a multidisciplinary approach associating expertise in cell culture, protein biochemistry, imaging technics (SEM, TEM, biphotonic microscopy, AFM, HS-AFM), analytical chemistry (PTR-MS and GC-FID) and chemoinformatics (QSPR approach). Moreover, MUFFIN is a fundamental research project, which will give the opportunity to a young researcher to develop his own research thematic and lead his own research group.


ANR Programme: Enjeux agroalimentaires (DS0502) 2014

Project ID: ANR-14-CE20-0001

Project coordinator:
Monsieur Francis Canon (Centre des Sciences du Goût et de l'Alimentation)

Project web site:


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The project coordinator is the author of this abstract and is therefore responsible for the content of the summary. The ANR disclaims all responsibility in connection with its content.